Sarisky Robert T, Crosson Paul, Cano Rachel, Quail Matthew R, Nguyen Tammy T, Wittrock Robert J, Bacon Teresa H, Sacks Stephen L, Caspers-Velu Laure, Hodinka Richard L, Leary Jeffry J
Department of Host Defense, The Antimicrobial and Host Defense Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, 1250 South Collegeville Road, UP 1450, Collegeville, PA 19426-0989, USA.
J Clin Virol. 2002 Jan;23(3):191-200. doi: 10.1016/s1386-6532(01)00221-9.
A number of in vitro assays are used to determine susceptibility of HSV to antiviral agents, but results from these in vitro assays do not necessarily correlate with treatment outcome.
A method with improved capability for identifying an isolate as acyclovir (ACV) or penciclovir (PCV) resistant when resistance is borderline could greatly improve the management of HSV disease.
A comparative evaluation of four in vitro assays, plaque reduction (PRA), DNA hybridization, plating efficiency (PEA) and plaque autoradiography (PAR) was performed to accurately identify and measure resistance of a TK-altered clinical HSV isolate (HSV-1 N4) from a patient who was non-responsive to ACV treatment. Two established criteria for the prediction of antiviral resistance, IC(50)> or =2.0 microg/ml or an IC(50) greater than 10x above a sensitive virus IC(50), as well as testing in human (MRC-5) and nonhuman (Vero and CV-1 monkey kidney) cell lines were evaluated.
The PRA and DNA hybridization assays accurately identified HSV-1 N4 as ACV(r) in human cells when using the 10x above sensitive virus IC(50) resistance criterion. Moreover, the PEA and PAR assays failed to classify HSV-1 N4 as drug resistant and indicate that these technologies alone are inadequate for identifying resistant virus.
The data presented herein indicate that the PRA and DNA hybridization assays most accurately identified an otherwise borderline-resistant isolate as drug resistant: (i) when a sensitive virus is used within each individual assay as a control, (ii) when ACV and PCV susceptibility is evaluated in human cells, and (iii) when the 10x above sensitive IC(50) criterion is used to classify a virus as drug-resistant. Testing of additional clinical samples is warranted to further confirm these findings.
多种体外试验用于确定单纯疱疹病毒（HSV）对抗病毒药物的敏感性，但这些体外试验的结果不一定与治疗结果相关。
当耐药性处于临界状态时，一种能够更准确地鉴定分离株对阿昔洛韦（ACV）或喷昔洛韦（PCV）耐药的方法，可极大地改善HSV疾病的治疗管理。
对四种体外试验进行了比较评估，即蚀斑减少试验（PRA）、DNA杂交试验、平板接种效率试验（PEA）和蚀斑放射自显影试验（PAR），以准确鉴定和测量来自一名对ACV治疗无反应患者的TK基因改变的临床HSV分离株（HSV-1 N4）的耐药性。评估了两种预测抗病毒耐药性的既定标准，即IC50≥2.0μg/ml或IC50比敏感病毒的IC50高10倍以上，以及在人（MRC-5）和非人（Vero和CV-1猴肾）细胞系中进行检测。
当使用比敏感病毒IC50高10倍的耐药标准时，PRA和DNA杂交试验在人细胞中准确地将HSV-1 N4鉴定为ACV耐药株。此外，PEA和PAR试验未能将HSV-1 N4分类为耐药株，表明仅这些技术不足以鉴定耐药病毒。
本文提供的数据表明，PRA和DNA杂交试验最准确地将原本临界耐药的分离株鉴定为耐药株：（i）在每个单独试验中使用敏感病毒作为对照时；（ii）在人细胞中评估ACV和PCV敏感性时；（iii）当使用比敏感IC50高10倍的标准将病毒分类为耐药株时。有必要对更多临床样本进行检测以进一步证实这些发现。
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